Identification of Ser465 as a novel PINK1 autophosphorylation site.
Identification of Ser465 as a novel PINK1 autophosphorylation site.
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Ser465 鉴定为新型 PINK1 自磷酸化位点
DOI：
10.1186/s40035-017-0103-7
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发表时间：
2017
影响因子：
12.6
通讯作者：
Tang BS
中科院分区：
文献类型：
--
作者：
Guo JF;Yao LY;Sun QY;Cui YT;Yang Y;Xu Q;Yan XX;Tang BS
PINK1基因是一种早发性常染色体隐性遗传性神经退行性疾病，是隐性家族性帕金森病6型(PARK6)的致病基因。据报道，PINK1同时具有自磷酸化和磷酸化活性，影响应激状态下的细胞损伤和其他生理反应。然而，关于PINK1自磷酸化位点的鉴定及其生理功能的研究尚未见报道。(1)采用质谱法对PINK1的自磷酸化位点进行鉴定，并通过放射自显影进一步证实这一结果。(2)用酶活性测定法比较Ser465突变体PINK1和致病突变体PINK1的活性。(3)我们使用脉冲追赶分析来检测Ser465是否会影响PINK1的降解。(4)免疫细胞化学染色在亚细胞水平上研究PINK1的亚细胞定位和Parkin转变。在我们的研究中，我们确定第465个丝氨酸残基(Ser465)是PINK1蛋白中的一个自动磷酸化位点。Ser465的失活可以降低PINK1的活性。PINK1 Ser465位点磷酸化的消散或过度都可以减缓其降解。PINK1的自动磷酸化有助于Parkin转运到线粒体，但对其亚细胞定位没有影响。PARK6原因突变T313M和R492X表现出与Ser465A突变PINK1蛋白相同的特征，如降低PINK1激酶活性并影响其与Parkin的相互作用。Ser465是PINK1的自磷酸化位点之一，影响PINK1的活性。此外，Ser465还参与PINK1的降解和Parkin向线粒体的转运。Thr313和Arg492上的两个新的PARK6突变T313、M和R492X与Ser465突变相似，包括PINK1磷酸化活性降低和Parkin亚细胞定位。
PINK1 (PTEN-induced putative kinase 1) gene is the causal gene for recessive familial type 6 of Parkinson’s disease (PARK6), which is an early-onset autosomal recessive inherited neurodegenerative disease. PINK1 has been reported to exert both autophosphorylation and phosphorylation activity, affecting cell damage under stress and other physiological responses. However, there has been no report on the identification of PINK1 autophosphorylation sites and their physiological functions. (1) We adopted mass spectrometry assay to identify the autophosphorylation site of PINK1, and autoradiography assay was further conducted to confirm this result. (2) Kinase activity assay was used to compare the kinase activity of both Ser465 mutant PINK1 and disease-causing mutant PINK1. (3) We use Pulse-chase analysis to measure whether Ser465 may affect PINK1 degradation. (4) Immunocytochemistry staining was used to study the PINK1 subcellular localization and Parkin transition in subcellular level. In our study, we identified the 465th serine residue (Ser465) as one of the autophosphorylation sites in PINK1 protein. The inactivation of Ser465 can decrease the kinase activity of PINK1. Either dissipated or excessive Ser465 site phosphorylation of PINK1 can slow down its degradation. PINK1 autophosphorylation contributes to the transit of Parkin to mitochondria, and has no effect on its subcellular localization. PARK6 causal mutations, T313 M and R492X, display the same characteristics as Ser465A mutation PINK1 protein, such as decreasing PINK1 kinase activity and affecting its interaction with Parkin. Ser465 was identified as one of the autophosphorylation sites of PINK1, which affected PINK1 kinase activity. In addition, Ser465 is involved in the degradation of PINK1 and the transit of Parkin to mitochondria. T313 M and R492X, two novel PARK6 mutations on Thr313 and Arg492, were similar to Ser465 mutation, including decreasing PINK1 phosphorylation activity and Parkin subcellular localization.
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影响因子：
9.8
作者：
Pridgeon JW;Olzmann JA;Chin LS;Li L
通讯作者：
Li L
影响因子：
168.9
作者：
Kalia, Lorraine V.;Lang, Anthony E.
通讯作者：
Lang, Anthony E.
影响因子：
3.5
作者：
Silvestri, L;Caputo, V;Casari, G
通讯作者：
Casari, G
影响因子：
4.8
作者：
Matenia, Dorthe;Hempp, Cindy;Mandelkow, Eva-Maria
通讯作者：
Mandelkow, Eva-Maria
DOI：
10.1073/pnas.0500617102
发表时间：
2005-04-19
影响因子：
11.1
作者：
Beilina, A;Van Der Brug, M;Cookson, MR
通讯作者：
Cookson, MR